Number Of Positive Samples Research Articles

Viral mapping in COVID-19 deceased in the Augsburg autopsy series of the first wave: A multiorgan and multimethodological approach.

COVID-19 is only partly understood, and the level of evidence available in terms of pathophysiology, epidemiology, therapy, and long-term outcome remains limited. During the early phase of the pandemic, it was necessary to effectively investigate all aspects of this new disease. Autopsy can be a valuable procedure to investigate the internal organs with special techniques to obtain information on the disease, especially the distribution and type of organ involvement. During the first wave of COVID-19 in Germany, autopsies of 19 deceased patients were performed. Besides gross examination, the organs were analyzed with standard histology and polymerase-chain-reaction for SARS-CoV-2. Polymerase chain reaction positive localizations were further analyzed with immunohistochemistry and RNA-in situ hybridization for SARS-CoV-2. Eighteen of 19 patients were found to have died due to COVID-19. Clinically relevant histological changes were only observed in the lungs. Diffuse alveolar damage in considerably different degrees was noted in 18 cases. Other organs, including the central nervous system, did not show specific micromorphological alterations. In terms of SARS-CoV-2 detection, the focus remains on the upper airways and lungs. This is true for both the number of positive samples and the viral load. A highly significant inverse correlation between the stage of diffuse alveolar damage and viral load was found on a case and a sample basis. Mediastinal lymph nodes and fat were also affected by the virus at high frequencies. By contrast, other organs rarely exhibited a viral infection. Moderate to strong correlations between the methods for detecting SARS-CoV-2 were observed for the lungs and for other organs. The lung is the most affected organ in gross examination, histology and polymerase chain reaction. SARS-CoV-2 detection in other organs did not reveal relevant or specific histological changes. Moreover, we did not find CNS involvement.

Gut microbiota and urinary tract infections in children

Aim. To evaluate the role of intestinal dysbiosis in the development of urinary tract infections in children admitted to a multidisciplinary hospital.Materials and Methods. We performed a bacteriological analysis of 2,694 urine samples collected from ≤ 3-years-old children who have been admitted to a multidisciplinary hospital (Kemerovo, Russian Federation). Urine specimens were inoculated by the 4-sector technique. Concurrently, we quantified the intestinal microbiota and its antimicrobial resistance in 100 children with urinary tract infections.Results. Titers of pathogenic microbes significantly differed in patients from distinct units (p = 0.02). The highest number of positive samples was detected in the neonatal pathology and neonatal intensive care units (43.8% each) as well as urology unit (37.9%). The most frequent pathogens belonged to Enterobacteriaceae family, in particular Escherichia coli, Klebsiella spp., Enterobacter spp., and Proteus spp. All studied children (100/100) suffered from intestinal dysbiosis. Titers of Bifidobacterium spp. and Lactobacillus spp. were reduced to 6 (5.0; 8.0) and 5 (4.0; 6.0) lg CFU/g, respectively. In 89.2% children, Escherichia coli lac+ levels were elevated to 9-10 lg CFU/g. Of note, 18.9% children had high Escherichia coli lac- titers [8 (6.0; 9.0) lg] and 24.3% had high Escherichia coli hly+ titers [5 (4.5; 6.0) lg] in the intestine. We have also found a high frequency (44.6%) and density [8.1 (7.0; 8.5) lg] of Klebsiella spp., in the intestinal mucosa of such patients. Notably, Enterobacter spp. and Proteus spp. were abundant [7 (5.0; 8.0) CFU/g] in the intestinal microbiota of 10.8% children. The prevalence of resistance strains in the studied setting reached 63%.Conclusions. More than 75% children with urinary tract infections suffer from intestinal dysbiosis. The microbiome of these patients was predominantly composed of Enterobacteriaceae and was characterized by high titers of Escherichia coli lac+, Klebsiella spp., Enterobacte spp., and Proteus spp. Similar profile of antimicrobial resistance in urinary and intestinal isolates of enterobacteria suggests intestinal microbiome as the main source of pathogens causing urinary tract infections in children.

Seasonal dynamics of gastrointestinal strongylatosis in the European bison

The purpose of the research is studying seasonal dynamics of gastrointestinal strongylatosis in bison in the Central Region of the Russian Federation.Materials and methods. The bison of different age and sex was studied in the Prioksko-Terrasny Nature Reserve, the Moscow Region. Fresh feces collected near feed troughs in enclosures in different seasons for three years were examined according to generally accepted methods. To analyze the infection rate of gastrointestinal strongylates in the European bison by months throughout the year, we took average monthly temperature and humidity values for 2018, 2019, and 2020 into account.Results and discussion. The maximum number of positive samples was diagnosed in the warm season of the year, namely, in summer and in the first months of autumn: August, September and October 2018 (79.3, 83.3 and 82.9%), August, October, November and December 2019 (73.7, 76.9, 77.1 and 77.8%), and June, July, August and October 2020 (85.6, 87.2, 88.0 and 86.3%). Ambient temperatures and precipitations affect significantly on the time of larvae development in the environment, which can be clearly seen in 2019.

The influence of oral antimicrobial peptide content on the quantitative microflora composition in periodontal pockets among residents of a large industrial region

The aim was to study the influence of periodontal pocket microecology state on the local nonspecific resistance in steelworkers with generalized periodontitis. Materials and methods. In total, 178 patients were examined. The study group consisted of 126 patients with generalized periodontitis of initial (n = 8), I (n = 32), II (n = 68) and III (n = 18) degree of severity, chronic course, exposed to work-related hazardous agents. The comparison group consisted of 32 patients with periodontitis of initial (n = 5), I (n = 10), II (n = 11) and III (n = 6) degree of severity without exposure to harmful conditions of steel industry. The control group included 20 otherwise healthy individuals. Detection of the main five periodontopathogenic microorganisms in the crevicular fluid was carried out by the polymerase chain reaction method. The levels of lactoferrin and cathelicidin LL-37 were measured by the enzyme-linked immunosorbent assay method. Results. Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis were less common, than other opportunistic bacteria in both study groups with initial severity of generalized periodontitis. With the disease development, there was a tendency to increase the number of positive samples for all microorganisms. A correlation was found between the microorganism presence in the sample and the severity of periodontal disease. In the patients of clinical groups, the concentration of cathelicidin LL-37 in the oral fluid was reduced. In the study group, a moderate inverse correlation between cathelicidin and Porphyromonas gingivalis as well as a weak inverse correlation between cathelicidin and Prevotella intermedia were revealed. Lactoferrin was correlated directly and moderately with Porphyromonas gingivalis and Prevotella intermedia but weakly - with Treponema denticola. In the comparison group, there were a statistically significant moderate inverse correlation between cathelicidin and Porphyromonas gingivalis as well a direct correlation between lactoferrin and Porphyromonas gingivalis. Correlations between the concentrations of cathelicidin and lactoferrin in the oral fluid and other periodontopathogens were revealed to be weak. Conclusions. The studies have shown significant disruptions of the oral cavity microecology in the patients exposed to harmful effects of steel industry. The correlation between the concentration of antimicrobial peptides and periodontal pathogens in the periodontal pockets has been found.

Evaluation of Anti-nuclear Antibodies (ANA) Patterns for aging population in Hong Kong

Abstract Objective Our aim was to investigate the association among aging population, sex and Antinuclear Antibody (ANA) patterns expressed in Hong Kong population for improving the autoimmune diagnosis by using ANA HEp-2 slides under fluorescent microscope. Methods A total of 50 blood samples from aging population in both males and females were collected and examined under fluorescent microscope. In this research, a HEp-2 complete Kits (Bio-Rad Kallestad) with the methodology of indirect fluorescent antibody (IFA). Positive ANA patterns classified in International Consensus on ANA Patterns (ICAP) are distinguished. Results In this experiment, we obtained that the number of positive samples was 43 (86%), the number of suspected (weak) positive samples was 9 (21%) and the number of negative samples was 7 (14%). In 43 positive samples, 24 out of 27 females (88.9%) and 19 out of 23 males (82.6%) were aged between 70 – 91. 21 in 43 positive samples in the result were within age group 70–78 (48.8%). 17 in 43 positive samples in the result were within age group 80–88 (39.5%). 5 in 43 positive samples in the result were within age group 90–92 (11.6%). The most frequent positive ANA patterns included AC10 – (7 patients, median age = 81, 20%), AC19 – (6 patients, median age = 82, 17%), and AC7 – Few Nuclear Dots (5 patients, median age = 84, 15%). We found that ANA positivity rate was significantly higher in female than the male in Hong Kong (6.3%). Conclusion Our data indicated that female aged 70–78 is more likely to result higher positivity rate of ANA than male. According to data of our study, this significantly supports the further study of the trend in the aging population who are easier to have an ANA related autoimmune disease to take better care for elderly in Hong Kong.

Evaluation of the GENE-UP® EHEC Detection Method for the Detection of Enterohemorrhagic E. coli in Select Foods: Collaborative Study: First Action Method 2020.06.

The GENE-UP® EHEC assay (Performance Tested MethodSM 121806) is a real-time PCR molecular detection method that utilizes fluorescence resonance energy transfer proprietary hybridization probes for the rapid detection of enterohemorrhagic Escherichia coli (EHEC) in select foods. The purpose of this validation was to evaluate the method's interlaboratory performance and submit the results to AOAC INTERNATIONAL for adoption as a First Action Official Method of AnalysisSM for the detection of EHEC in select foods. The GENE-UP method was evaluated in a multi-laboratory study as part of the MicroVal VALIDATION certification process using unpaired test portions for one food matrix, raw ground beef (85% lean). Collaborators evaluated the candidate method using either an automated or manual lysis procedure. The candidate method was compared to the ISO/TS 13136:2012 method. Data from 17 participants from 15 laboratories throughout the European Union were evaluated. Three levels of contamination were evaluated: a non-inoculated control level (0 CFU/test portion), a low contamination level (∼1 CFU/test portion), and a high contamination level (∼10 CFU/test portion). Data from the study were analyzed according to the probability of detection (POD) statistical model. The difference in laboratory probability of detection (dLPODC) values with 95% confidence interval between the candidate and reference method results were -0.01 (-0.04, 0.02), 0.23 (0.07, 0.39), and 0.06 (0.01, 0.12) for the non-inoculated, low, and high contamination levels, respectively. For the candidate method, values obtained for repeatability and reproducibility were similar to the reference method and indicated minimal variation between samples or between laboratories. No discrepant results (false positive or false negative) were observed for each contamination. A statistical difference was calculated between the candidate and reference method at the low and high inoculation levels, with the candidate method detecting a higher number of positive samples indicating a higher sensitivity than the reference method. No differences in the recovery of the target analyte were observed between the manual and automated lysis procedures. The GENE-UP EHEC Detection Method provides end users a rapid, easy-to-use workflow for the detection of EHEC in food matrixes.

Molecular epidemiology of Cryptosporidium spp. in an agricultural area of northern Vietnam: A community survey.

The purpose of this study was to investigate the occurrence of Cryptosporidium infection and the potential for transmission of Cryptosporidium spp. between animals and humans in northern Vietnam. A total of 2715 samples (2120 human diarrheal samples, 471 human non-diarrheal samples, and 124 animal stool samples) were collected through our community survey in an agricultural area. All samples were tested for Cryptosporidium spp. by direct immunofluorescence assay (DFA) using a fluorescent microscope. DNA extraction, PCR amplification of three genes (COWP, SSU-rRNA, and GP60), and sequencing analysis were performed to identify Cryptosporidium spp. Of 2715 samples, 15 samples (10 diarrheal samples, 2 non-diarrheal samples, and 3 animal stool samples) tested positive by PCR for the COWP gene. Three species of Cryptosporidium spp. were identified as C. canis (from six human diarrheal samples, two human non-diarrheal samples, and one dog sample), C. hominis (from four human diarrheal samples), and C. suis (from two pig samples) by sequencing the amplified COWP and/or SSU-rRNA genes. In terms of C. hominis, the GP60 subtype IeA12G3T3 was detected in all four human diarrheal samples. Although the number of positive samples was very small, our epidemiological data showed that the emerging pattern of each of the three species (C. canis, C. hominis, and C. suis) was different at this study site. While C. hominis and C. suis were only detected in human and pig samples, respectively, C. canis was detected in samples from both dogs and humans. We suspect that C. canis infections in humans at this study site may be due to environmental contamination with animal and human feces.

Chemical residues in emulsion type meat products (Frankfurter & Luncheon)

In this study total of 150 samples of emulsion type meat products (Luncheon & Frankfurter) (75 of each) were collected from local and supermarkets in Giza governorate, Egypt. Samples were subjected to determination of residue of antibiotics, zeranol, trenbolone, heavy metals (Cadmium, lead, Zinc and Arsenic) and pesticides (OPPs & OCPs). The study proved that the incidence percent of antibiotics, organophosphorus pesticides and arsenic were zero in all samples, zeranol percent in frankfurter was 0% while is detected in (25) samples of luncheon with mean (0.0732 ppb). Concerning trenbolone in luncheon and frankfurter with the same number of positive samples (50) for each with mean (0.2213) and (0.1776) ppb respectively. Detected residues of lead were present in 50 samples of luncheon and frankfurter with mean (0.0504) and (0.0165)ppm respectively. The cadmium was recorded in 50 samples of frankfurter with mean (0.0044 ppm), while were reported in luncheon with mean (0.0019 ppm). The zinc was recorded in (75 samples) of frankfurter with mean (0.597 ppm), while it was recorded in 50 samples of luncheon with mean (0.3839 ppm). The PP-DDT was recorded with highest level in luncheon with value (3.638 ppm) while PP.DDT with lower level was reported in frankfurter with value (1.69856 ppm). Methoxychlor was recorded only in luncheon with value (59.02398 ppm), but not detected in frankfurter. Aldrin was detected only in frankfurter with value (0.756 ppm). Endrin was not recorded in luncheon and frankfurter. PP. DDD was recoded in luncheon with value (0.445 ppm) but not in frankfurter.

Diagnosis of COVID-19 Using Machine Learning and Deep Learning: A Review

Background: This paper provides a systematic review of the application of Artificial Intelligence (AI) in the form of Machine Learning (ML) and Deep Learning (DL) techniques in fighting against the effects of novel coronavirus disease (COVID-19). Objective & Methods: The objective is to perform a scoping review on AI for COVID-19 using preferred reporting items of systematic reviews and meta-analysis (PRISMA) guidelines. A literature search was performed for relevant studies published from 1 January 2020 till 27 March 2021. Out of 4050 research papers available in reputed publishers, a full-text review of 440 articles was done based on the keywords of AI, COVID-19, ML, forecasting, DL, X-ray, and Computed Tomography (CT). Finally, 52 articles were included in the result synthesis of this paper. As part of the review, different ML regression methods were reviewed first in predicting the number of confirmed and death cases. Secondly, a comprehensive survey was carried out on the use of ML in classifying COVID-19 patients. Thirdly, different datasets on medical imaging were compared in terms of the number of images, number of positive samples and number of classes in the datasets. The different stages of the diagnosis, including preprocessing, segmentation and feature extraction were also reviewed. Fourthly, the performance results of different research papers were compared to evaluate the effectiveness of DL methods on different datasets. Results: Results show that residual neural network (ResNet-18) and densely connected convolutional network (DenseNet 169) exhibit excellent classification accuracy for X-ray images, while DenseNet-201 has the maximum accuracy in classifying CT scan images. This indicates that ML and DL are useful tools in assisting researchers and medical professionals in predicting, screening and detecting COVID-19.

Effectiveness of an institutional COVID-19 central sampling team during pandemic at a tertiary care centre.

Background:An efficient sampling is one of the key methods to identify all those affected by coronavirus disease 2019 (COVID-19).Objectives:To analyze how efficient setting up of a central sampling team would be to prevent any outbreak within the institution by minimizing the movement of suspected COVID-19 patients admitted in the inpatient wards. The secondary objective was to train maximum resident doctors to collect samples of admitted patients.Methodology:A central sampling team comprising of resident doctors from various departments was made who did sampling of the suspected COVID-19 inpatients admitted under various specialties.Results:There were a total of 341 patients [209 males (61.29%), 132 females (38.7%)] and 335 patients underwent sampling. There was a positive correlation between: (1) number of calls from a department vs percentage of positive samples in that department [Pearson correlation coefficient (R) = 0.47; P = 0.026], (2) number of samples taken by resident of a particular department from central sampling team vs number of positive samples taken by resident of that department [R = 0.8739, P = 0.01] and (3) number of visits to a department vs number of residents trained in that department [R = 0.93; P = 0.00001].Conclusion:Formulation of a central sampling team led to changes like a separate donning and doffing area in each ward and training of many resident doctors posted in different wards. This made each ward self-sufficient in collection of samples. This venture also ensured minimal movement of suspected COVID-19 patients in the hospital and thus least exposure to the hospital staff.

417. Comparative Assessment of Multiple SARS-CoV-2 Antibody and Neutralization Assays from Blood Samples in COVID-19 Infected Patients

BackgroundThe severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, COVID-19) has caused a world-wide pandemic. Diagnosis is usually made by an RT-PCR test from a respiratory sample. A number of tests are available for antibody detection or assessment, including rapid, enzyme immunoassays (EIA) and neutralization. However, characterization of the antibody immune response is not well documented and the clinical significance of COVID antibodies remains largely unknown. In addition, comparison of results across different assay formats using identical samples has not been rigorously studied, making clinical interpretation of serologic tests difficult.Assessment of multiple SARS-CoV-2 antibody and neutralization assays from blood samples in COVID-19 infected patients Methods1–5 serial (total 33) serum or plasma samples from 14 patients who were positive for SARS-CoV-2 by EUA authorized RT-PCR assays from nasopharyngeal specimens where tested with the following COVID-19 antibody tests: LFA rapid tests (Chembio DPP IgM/IgG, SD Biosensor Standard IgM/IgG, BTNX Rapid Response IgM/IgG), and EIA tests (BioRad Platelia SARS-CoV-2 Total antibody-IgG/IgM/IgA, EuroImmun SARS-CoV-2 IgG, and EuroImmun SARS-CoV-2 IgA). See Table 1 for results and EUA. Results were recorded as positive, negative, or equivocal. Additionally, antibody neutralization was assessed on matched samples.ResultsMean age of SARS-CoV-2 positive patients was 73 years (range 65–89), 11/14 had symptoms, all were male and hospitalized (6 ICU), and 3 died. Average number of days serum was collected after RT-PCR positivity was 13.5 days (range -3 to 46 d). BTNX assay was only tested on 16 samples. Among all assays, total concordance of results was 91%. When only IgG/IgM or total antibody assays were considered, concordance of results was 96% (Table). IgA specific results were discordant in 9/33 (27%) of samples compared to other assays. Two patients were negative in all assays in serial samples collected within one week of PCR positivity. Antibody neutralization was detected, but not from all samples.ConclusionIn general, there was good agreement among antibody detection assays. Neutralization may reflect disease outcome. The study was limited by the number of positive samples and patient number, and at the time specificity was not addressed for all the assays.Disclosures All Authors: No reported disclosures

LB-15. A Trans-Governmental Collaborative Effort to Independently Evaluate SARS-CoV-2 Serology Assays Using Well-Characterized Sample Panels

BackgroundThe emergence of the novel coronavirus, SARS-CoV-2, created a crucial need for accurate tests for diagnosis, assessment of prior infection, and understanding its natural history. Serology assays play an important role in the assessment of anti-viral immune responses and previous infections. Evaluation of serology assays with well-characterized serum and/or plasma samples is critical to determine assay performance. CDC, FDA and NCI’s Frederick National Laboratory for Cancer Research (NCI-FNLCR) have established a collaborative network to independently evaluate commercial antibody tests prior to their authorization.MethodsPositive (n=30) serum samples with a range of anti-SARS-CoV-2 antibody titers (Table) and negative (n=80) serum and/or plasma samples were selected to establish performance evaluation panels (PEVs). Three PEVs with similar overall antibody titer distribution have been created. Negative samples were collected prior to 2020, before the SARS-CoV-2 pandemic. Positive samples were from patients previously confirmed to have SARS-CoV-2 using a nucleic acid amplification test. Each sample was characterized at CDC and NCI-FNLCR for presence/absence of SARS-CoV-2 IgM and IgG antibodies using a SARS-CoV-2 spike enzyme linked immunosorbent assay (ELISA). NCI-FNLCR also performed a SARS-CoV-2 spike Receptor Binding Domain (RBD) IgG ELISA. Positive samples were assessed at multiple dilutions. Manufacturers submitted their serology assays for evaluation by this program. The sensitivity of each test was assessed for each antibody class (IgG and IgM) and in a combined manner, where a positive result for either antibody was considered as a positive result. For combined specificity, a negative result meant a sample was negative for both antibodies (IgG and IgM).Number of positive samples with anti-SARS-CoV-2 spike antibodies for each panel (n=30) ResultsTo date, 53 serology assays have been evaluated. Sensitivity ranged from 30.0% to 100% for IgG, from 10.0% to 100% for IgM, and the combined specificity ranged from 57.5% to 100%. For 2 assays that measure total Ig, sensitivity was 96.7% and 100%.ConclusionThis program completed over 50 performance evaluations with well-characterized PEVs. Results have been used to inform FDA regulatory decisions and are publicly available on FDA’s website.Disclosures Cristina Cassetti, PhD, Nothing to disclose

The reliability of SARS-CoV-2 IgG antibody testing – a pilot study in asymptomatic health care workers in a Croatian university hospital

AimTo evaluate three fully automated serological assays in terms of reactivity to SARS-CoV-2 immunoglobulin G (IgG) and perform SARS-CoV-2 IgG antibody testing among asymptomatic health care workers (HCW) at the University Hospital Center Zagreb.MethodsThree IgG serological assays (Abbott SARS-CoV-2 IgG, Elecsys Anti-SARS-CoV-2, and MAGLUMI 2019-nCoV IgG) were initially evaluated by analyzing 42 samples from confirmed COVID-19-recovered patients and 48 negative individuals. A total of 1678 HCW ( ~ 30% of all hospital employees) were screened for SARS-CoV-2 IgG with the Abbott assay, run on Abbott Architect i2000SR. The samples exceeding the predefined cut-off (1.4 S/C) were reanalyzed with the Elecsys, MAGLUMI, and VIDAS SARS-COV-2 IgG assays.ResultsInitially, the MAGLUMI 2019-nCoV IgG produced 26.2% false negatives and the Elecsys Anti-SARS-CoV-2 produced one false positive. Among 1678 HCW, the Abbott assay showed only 10 (0.6%) positive results, with mostly mildly elevated signals. Nine of these samples were non-reactive when they were retested with the Elecsys, MAGLUMI, and VIDAS assays. As for the one remaining sample, it was positive when tested with the Elecsys assay, while the other two assays yielded negative results.ConclusionsSARS-CoV-2 IgG seroprevalence among asymptomatic HCW in our hospital setting was low, with different assays indicating a different number of positive samples. One of the assays yielded a large false negative rate. These findings can be attributed to differences in assay formulation but also to heterogeneity and diverse reactivity of antibodies against SARS-CoV-2 antigens.

Learn to Extract Building Outline from Misaligned Annotation through Nearest Feature Selector

For efficient building outline extraction, many algorithms, including unsupervised or supervised, have been proposed over the past decades. In recent years, due to the rapid development of the convolutional neural networks, especially fully convolutional networks, building extraction is treated as a semantic segmentation task that deals with the extremely biased positive pixels. The state-of-the-art methods, either through direct or indirect approaches, are mainly focused on better network design. The shifts and rotations, which are coarsely presented in manually created annotations, have long been ignored. Due to the limited number of positive samples, the misalignment will significantly reduce the correctness of pixel-to-pixel loss that might lead to a gradient explosion. To overcome this, we propose a nearest feature selector (NFS) to dynamically re-align the prediction and slightly misaligned annotations. The NFS can be seamlessly appended to existing loss functions and prevent misleading by the errors or misalignment of annotations. Experiments on a large scale aerial image dataset with centered buildings and corresponding building outlines indicate that the additional NFS brings higher performance when compared to existing naive loss functions. In the classic L1 loss, the addition of NFS gains increments of 8.8% of f1-score, 8.9% of kappa coefficient, and 9.8% of Jaccard index, respectively.

PREVALENCE OF SMALL RUMINANT LENTIVIRUS INFECTIONS IN SHEEP AND GOATS IN SOME REGIONS OF VOJVODINA PROVINCE

Small Ruminant Lentivirus causes a chronic lifelong, multisystemic diseases in sheep and goats with or without clinical manifestation. Maedi-visna virus (MVV) and Caprine arthritis encephalitis virus (CAEV) are often considered together as small ruminants lentivirus (SRLV) due to their phylogenetic correlation and the interspecies transmission between sheep and goats.
 During the period of four years, from 2015-2018, an annual monitoring was done on male animals (rams and bucks) used for breeding, for determination of antibodies against SRLV with ELISA serological method. In total, serum samples from 6732 animals were analyzed.
 During the study period, seroprevalence for small ruminant lentivirus has rised from 4,17% to 16,21%, with an overall value of 12% for the whole period. Over the years seroprevalence has been changing with increasing tendency. The highest seroprevalence in rams was in 2018, when the largest number of samples was examined. In bucks, the highest number of positive samples was found in 2015. From 2014 until 2018, the overall seroprevalence in rams and bucks has increased significantly. From the study results it can be seen that annual monitoring program is very important and has to be held in the future in order to have the disease under control. The program helps in rising the awareness and knowledge of the owners about SRLV and the importance of breeding seronegative animals.

Prevalence of Salmonella enterica subsp. diarizonae serotype 61:k:1:5:(7) in nasal secretions and stool of sheep flocks with and without cases of chronic proliferative rhinitis

Salmonella enterica subsp. diarizonae serotype 61: k: 1,5, (7) (SED) is a microorganism well adapted to sheep; however, it has also been described producing chronic proliferative rhinitis (CPR) in ovine. CPR causes a proliferative inflammation of the ventral nasal turbinates that may totally obstruct the nasal cavity.The main objective of the present study was to investigate the prevalence of SED in nostrils and stool of sheep without CPR clinical signs in commercial sheep farms of Spain with and without previous clinical cases of CPR.Five samplings were performed in 10 commercial sheep farms for one year. Samples from nostrils and faeces were taken from four animals without CPR visible clinical signs that belonged to four different age ranges at each farm visit.The prevalence of positive animals was 45.3 %, and the number of positive samples in nostrils was higher than in faeces (38.5 % vs 22.5 %). Only on one farm was no positive result obtained in the entire study. In almost all positive farms, sheep belonging to the youngest age ranges accounted for more than 50 % of positive isolates. Finally, farms with a previous diagnosis of CPR were 1.784 times more likely to have an animal with positive isolation than farms without a previous diagnosis. This could suggest that the infection pressure in the farm might favour the occurrence of clinical cases of the disease. However, further studies will be necessary to unravel why this saprophytic bacterium is able to cross the epithelial barrier causing severe rhinitis in certain animals.

Expression of ANA patterns by Indirect Fluorescent Assay in a local Hong Kong population from private sector

Abstract Objective Our aim was to investigate the association among age, sex and Antinuclear Antibody (ANA) patterns expressed in Hong Kong population for improving the autoimmune diagnosis by using ANA HEp-2 slides under fluorescent microscope. Methods A total of 65 blood samples from both male and female were collected and examined under fluorescent microscope. Haemolysed samples were excluded. In this research, a HEp-2 complete Kits with the methodology of indirect fluorescent antibody (IFA) was utilized for the detection and semi - quantification of antinuclear antibodies to distinguish positive ANA patterns classified in International Consensus on ANA Patterns (ICAP) for human body. Results In this experiment, we obtained that the number of positive samples was 6 (9%), the number of suspected (weak) positive samples was 4 (6%) and the number of negative samples was 55 (85%). In 6 positive samples, they were all females aged between 59 – 89. 5 in 6 positive samples in the result were over age 69 (83%). The positive ANA patterns included AC2 – Speckled, AC3 – Centromere, AC6 – Discrete Nuclear Dots, AC7 – Discrete Nuclear Dots, and AC8 – Nucleolar patterns separately. No AC-29 - DNA topoisomerase I was examined and resulted. We found that Hong Kong have a higher distribution of ANA positive frequency for female (9.23%) than male. Conclusion Our data indicated that ANA patterns were found in female aged above 69 is more likely to suffer from autoimmune disease related to ANA, providing an important foundation for considering and studying the trend in the elderly who are easier to have an ANA related autoimmune disease in Hong Kong.

Occurrence of Mycoplasma gallisepticum in wild birds: A systematic review and meta-analysis.

Mycoplasma gallisepticum is one of the most important poultry pathogens that can also infect wild birds, but knowledge of potential non-poultry hosts that could be reservoirs of M. gallisepticum is limited. For the paper presented here, we screened three databases (PubMed, Scopus, and the Web of Knowledge) to find articles on the occurrence of M. gallisepticum in different wild bird species that were published between 1951 and 2018. Among 314 studies found, we selected and included 50 original articles that met the pre-established criteria. From those publications we extracted the following information: name of the first author, year of publication, year of sample isolation, country, region, number of birds sampled, number of birds tested by each method, number of positive samples, diagnostic criteria, and if birds were wild or captive. Because different detection techniques were used to confirm the presence of M. gallisepticum in one animal, we decided to perform the meta analyses separately for each method. The estimated prevalence of M. gallisepticum in wild birds was different by each method of detection. Our summary revealed that M. gallisepticum was present in 56 species of bird belonging to 11 different orders, of which 21 species were reported suffering both past and current infection. Our work provides information on wild bird species that could be considered potential reservoirs or carriers of M. gallisepticum and could be helpful to set the direction for future research on the spread and phylogeny of M. gallisepticum in different hosts.

Environmental contamination with cytotoxic drugs in 15 hospitals from 11 European countries—results of the MASHA project

Abstract Background: Evaluating environmental contamination with cytotoxic drugs in hospitals is one of the fundamental requirements to ensure the occupational safety of all healthcare professionals. The European Society of Oncology Pharmacy (ESOP) undertook the first independent, multicenter, pan-European study, involving over a dozen hospitals from 11 different countries, to measure the state of cytotoxic contamination in the workplace. Objective: To obtain an overview of the current situation in European hospitals with regards to cytotoxic drug contamination at various sites, including drug preparation (pharmacy) and administration areas (ward). The secondary objectives are to evaluate the environmental contamination with cytotoxic drugs circulating within a facility known as the hospital medication system (process flow of drug) and to evaluate the impact of changes in local cleaning practices. Materials and methods: The study was carried out at 15 hospitals in Europe evaluating the surface contamination in the preparation and administration areas before (part I) and after (part II and part III) training and the implementation of ESOP cleaning recommendations. Assessment of surface contamination with 11 antineoplastic drugs was performed using wipe samples taken from 10 comparable surfaces (5 each in the preparation and administration areas). These samples were analyzed by liquid chromatography–tandem mass spectrometry. Results: The study demonstrated the presence of surface contamination in preparation and administration areas in all hospitals, with measurable amounts of at least 1 agent detected on sampled surfaces. Before the implementation of the ESOP cleaning recommendations, 324 out of 1595 results were positive (20%). In 11 of 15 hospitals (73%), substances were detected which were not prepared or administrated in the sampling day. After implementation of the ESOP recommendations, only 14% of results were positive (226/1639). Sixty-nine percent of wards (9/13) improved or stayed at the same level in the number of positive samples. The floors on the wards were shown to be the most frequently contaminated (42% of samples were positive). The amount of contamination in the pharmacies was not correlated to the amount of chemotherapy prepared nor to the use of special devices such as closed-system transfer devices. Conclusion: The MASHA study provides an overview of the contamination levels with cytotoxic drugs in European hospitals. Upon implementation of ESOP cleaning recommendations, improvements could be seen, with a reduced number of positive-wipe samples and lower amounts of surface concentration detected. The study demonstrates that improving standard work procedures is able to substantially reduce contamination in the workplace.

Light-driven antimicrobial activities of vitamin K3 against Listeria monocytogenes, Escherichia coli O157:H7 and Salmonella Enteritidis

Due to the lack of thermal inactivation steps, the fresh produce industry is actively looking for new, environmentally friendly and effective non-thermal control strategies to eliminate foodborne pathogens on produce surfaces and to prevent cross-contamination. Photodynamic inactivation of bacteria has emerged as a novel alternative approach to inactivate microorganisms on fresh produce. In this study, the photo-sensitizing efficacy of vitamin K3 (VK3) under UV-A and simulated sunlight and their antimicrobial efficacy against common foodborne pathogens in broth and on lemon surfaces were investigated. The minimal bactericidal concentration (MBC) of VK3 against Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella Enteritidis PT30 in darkness was 625.0, 156.3, and 625.0 μg/ml, respectively. When treatments were carried out under light, the MBC values of VK3 (VK3 + light) were reduced in strain, light source, and exposure time dependent manners. The enhanced antimicrobial effect can be attributed to the production of reactive oxygen species of VK3, including hydroxyl radical and singlet oxygen under light exposure. Based on scanning electron microscope examination, VK3 + light treatment damaged the cell integrity of L. monocytogenes and Salmonella; and altered the cell morphology of E. coli O157:H7 including filament formation and fewer fimbriae/flagella. When applied on artificially inoculated lemon surfaces, VK3 + light treatments at MBC levels immediately reduced the levels of L. monocytogenes and E. coli O157:H7 by more than 5.0 Log10 CFU/disk and S. Enteritidis PT30 by ~4.0 Log10 CFU/disk. Bacterial populations of all tested strains were reduced to below the limit of detection (1.0 Log10 CFU/disk) after one day of storage at 4 °C; the numbers of positive samples after enrichment further decreased as the storage time increased, demonstrating the continuous antimicrobial effects the VK3 generated on lemon surfaces. In summary, the results of this study demonstrated the promising antimicrobial effects of VK3 + light treatments and their great application potential for fresh produce industry.